When I design guide RNA of CRISPR/CAS9 system, the gRNAs always offtarget to pseudogenes of my gene. These pseudogenes are unable to coding protein according to NCBI. Can I use these gRNAs to do gene knock-out?
If you can confirm that your pesudogenes don't encode functional mRNAs or proteins, you should be able to use your vectors for experiments. For instance, you could use QRT-PCR or northern blot to show that these pesudogenes have no expression in the tissue you analysing .
@ Guo Dongyang,
Just wondering, has you resolved the problem yet. I am interested to know. Thanks.
By the way, as lone as the sgRNA and Cas9 can also target to the 'normal' copy of the gene, then it should be fine. You can still get a null allele of the normal copy. Don't you think so?
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology